The efficacy of some vaccines is enhanced by the presence of adjuvants added to their formulations or, in the case of live attenuated or killed whole cell vaccines, because of their endogenous adjuvant activity. The immune system responds robustly to these endogenous adjuvants, which includes Pathogen Associated Molecular Patterns, which stimulate innate immune responses through Pattern Recognition Receptors, such as TOLL-like receptors (TLRs). The development of most vaccine adjuvants has occurred despite little understanding of their overall mechanisms of immune enhancement. We hypothesized that TLR-dependent adjuvant activities are mediated through TLR stimulation of antigen presenting cells (APCs), and each APC type may play a unique role in the immune-stimulating ability of these adjuvants, including effects on downstream T cell stimulation. We used a mouse model where TLR/MyD88 signaling is prevented in specific APC types, in vivo, using loxP/cre recombinase transgenic mice (B cells, dendritic cells and macrophages) to investigate its role in vaccine adjuvant activity. We found that intact MyD88 signaling is essential, separately, in all three APC types for optimal TLR-ligand based adjuvant (PorB, CpG, MPLA), but not for TLR-independent
(Alum, MF59) adjuvant activity. However, the immune responses were reduced to the greatest extent in mice with macrophage specific MyD88 deletion (Mac-MyD88-/-). We demonstrated that TLR-dependent adjuvants are potent inducers of germinal center (GCs) formation needed for an effective and robust immune response. Interestingly, GCs are nearly absent in Mac-MyD88-/- mice upon immunization with TLR-dependent adjuvants, but not with TLR-independent adjuvants. Further investigations revealed a significant impairment in T cell cytokines important for GC formation in Mac-MyD88-/- mice when immunized with TLR-dependent adjuvants. Through these studies we discovered that vaccine formulated with PorB/OVA induced a robust and diverse T cell response including highly functional OVA-CD4 and CD8 T cells. These CD8 T cells are protective and significantly reduced the bacterial burden and increased survival in a Listeria mouse infection model. Our findings reveal that PorB has broad adjuvant activity, signaling through all three APC types, inducing strong and diverse humoral and cellular responses. These insights will allow for a more intelligent use of adjuvants in future vaccine development.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/19058 |
| Date | 03 November 2016 |
| Creators | Mosaheb, Munir |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
| Rights | Attribution-NonCommercial-NoDerivatives 4.0 International, http://creativecommons.org/licenses/by-nc-nd/4.0/ |
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